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Investigation of ionic channel and its * cance in *

离子通道及其*癌症的研究*

基本信息

批准号：
11670568
负责人：
NAKAJIMA Toshiaki
金额：
$ 0.96万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670568/
关键词：
Bronchial smooth muscle endothelin-1 ionic currents Patch champ K^+ channel Anti-serces Non-selective cation channels Biorchial asthma Brachial Smooth muscle Anti-sanse Kir2.1 Endothelin-1 Patch-clamg Intracellulor Ca^<2+> ions /

项目摘要

1) Electro-physiological effects of endothelin-1 on human bronchial smooth muscle cells : We investigated the effects of endothelin-1 and histamine on electro-physiological effects of endothelin-1 on human bronchial smooth muscle cells, by means of the patch-clamp techniques and Ca^<2+> measurements using fura-2 AM, Both endothelin-1 and histamine first increased [Ca^<2+>]_1 by the release of Ca^<2+> from store sites, then fol1owed by the sustained Ca^<2+> influx. The [Ca^<2+>]_1 increase induced by endothelin-1 was mediated by both ET_A and ET_B receptors. The activation of Ca^<2+>-permeable nonselective cation channels (I_<NS>) was involved in the sustained rise in [Ca^<2+>]_1, where ET_A and ET_B receptors were involved. The EC_<50> value for endothelin-1 to activate I_<N,S-> was approximately 10 nM.In addition, presence of both ET_A and ET_B mRNA was confirmed by RT-PCR in these cells and immunostaining of ET receptors in human airway preparations.2) K^+ channels in human bronchial smooth muscle cells : We examined the properties of K^+ channels, by using the patch clamp techniques and the RT-PCR analysis of mRNA and antisense oligonucleotide methods. In these cells, we found the voltage-dependent K^+ channels (K_v) and the inwardly rectifying K^+ channels (Kir), which contribute to forming the resting membrane potential. The Kir channels was blocked by low doses of extracellular Ba^<2+> ions with a IC_<50> of approximately - 1.8μM.RT-PCR analysis of mRNA showed transcripts for Kir2.1. but not Kir1.1, 2.2, and 2.3. Treatment of cells with antisense oligonucleotides targeted to Kir2.1 resulted in a significant decrease in current density of the Kir current. These results demonstrate that the inward rectifier K^+ current (Kir) is present in human bronchial smooth muscle cells. and the Kir2.1 gene encodes the Kir channel protein in these cells.

1) 内皮素-1对人支气管平滑肌细胞的电生理效应：我们通过膜片钳技术和使用fura-2 AM的Ca^<2+>测量，研究了内皮素-1和组胺对内皮素-1对人支气管平滑肌细胞的电生理效应的影响，内皮素-1和组胺首先增加[Ca^<2+>]_1 通过从储存位点释放 Ca^<2+>，然后是持续的 Ca^<2+> 流入。内皮素-1诱导的[Ca 2+ ]_1增加是由ET_A和ET_B受体介导的。 Ca^<2+>-渗透性非选择性阳离子通道(I_<NS>)的激活参与了[Ca^<2+>]_1的持续升高，其中ET_A和ET_B受体参与其中。内皮素-1激活I_<N,S->的EC_<50>值约为10nM。此外，通过RT-PCR在这些细胞中和人气道制剂中ET受体的免疫染色证实了ET_A和ET_B mRNA的存在。2)人支气管平滑肌细胞中的K^+通道：我们通过使用膜片钳技术和RT-PCR检查了K^+通道的特性 mRNA和反义寡核苷酸分析方法。在这些细胞中，我们发现了电压依赖性 K^+ 通道 (K_v) 和内向整流 K^+ 通道 (Kir)，它们有助于形成静息膜电位。 Kir通道被低剂量的细胞外Ba^2+离子阻断，IC_50约为～1.8μM。mRNA的RT-PCR分析显示Kir2.1的转录物。但不是 Kir1.1、2.2 和 2.3。用针对 Kir2.1 的反义寡核苷酸处理细胞导致 Kir 电流的电流密度显着降低。这些结果表明，内向整流 K^+ 电流 (Kir) 存在于人支气管平滑肌细胞中。 Kir2.1 基因编码这些细胞中的 Kir 通道蛋白。